Detection of Circulating Prostate Cells during Radical Prostatectomy by Standardized PSMA RT-PCR: Association with Positive Lymph Nodes and High Malignant Grade

Bettina Schmidt, Aristotelis G. Anastasiadis, Hans Helge Seifert, Knut H. Franke, Mototsugu Oya, Rolf Ackermann

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Background: The purpose of this study was to establish a quantitative standardized RT-PCR for the detection of Prostate Specific Membrane Antigen (PSMA)-expressing circulating cells and to evaluate clinical relevance in patients undergoing radical retropubic prostatectomy (RRP) for clinically localized prostate cancer (PCa). Materials and Methods: An external standard molecule (PSMA MIMIC) was constructed for standardization of PSMA RT-PCR reactions. It has the same sequence as endogenous PSMA, except for a central 85 bp deletion, allowing the amplification of both targets simultaneously with nearly the same amplification characteristics as in a nested PCR. PSMA RT-PCR was performed from peripheral blood samples of 73 patients with clinically localized PCa, 4 with metastatic PCa, 27 with benign prostatic hyperplasia (BPH) and 27 controls. Pre, intra- and postoperative blood samples were tested for circulating PSMA expressing cells in 54 out of 73 patients with clinically localized PCa. We also tested intraoperative blood samples of 19 BPH patients treated by transurethral or open surgery. Results: Endogenous PSMA signals from PCa patients varied between 850 and 9900 transcript molecules, corresponding to 2-20 PSMA-expressing cells/ml blood. Standardized RT-PCR using the PSMA MIMIC molecule revealed a significant decrease of "false-positives" in cancer-free controls (p=0.004). Controls could clearly be distinguished from prostate cancer patients based on PSMA PCR positivity (p=0.003). Thirty-two % of patients with localized prostate cancer, 11% of BPH patients and 7% of healthy controls were positive in standardized assays compared to 48%, 30% and 27% without PSMA MIMIC, respectively. Preoperatively, a correlation with tumor stage (p=0.030), grade (p=0.035) and Gleason Score (p=0.03) could be demonstrated in clinically localized PCa patients. Dissemination of prostate cells during surgery occurred in 32% of the RRPs and 21% of BPH patients. Positive PCR signals from intraoperative blood samples correlated with positive lymph node status (p=0.007) and tumor grade (p=0.005). Postoperative positive results correlated with grade (p=0.012) and Gleason Score (p=0.035). Conclusion: Counseling the patient with clinically localized prostate cancer can be challenging. Surgery may be, in retrospect, inappropriate in a number of patients due to preoperative understaging. This newly constructed external standard allows quantitative detection of circulating prostate cells, and therefore may open new perspectives for PSMA RT-PCR techniques as diagnostic assays and tools for post-therapeutic follow-up.

Original languageEnglish
Pages (from-to)3991-3999
Number of pages9
JournalAnticancer Research
Volume23
Issue number5 A
Publication statusPublished - 2003 Sep

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Prostatectomy
Prostate
Lymph Nodes
Polymerase Chain Reaction
Prostatic Neoplasms
Prostatic Hyperplasia
Neoplasm Grading
human glutamate carboxypeptidase II
Neoplasms
Counseling
Blood Cells

Keywords

  • Prostate cancer
  • PSMA
  • Radical prostatectomy
  • RT-PCR
  • Standardization

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Detection of Circulating Prostate Cells during Radical Prostatectomy by Standardized PSMA RT-PCR : Association with Positive Lymph Nodes and High Malignant Grade. / Schmidt, Bettina; Anastasiadis, Aristotelis G.; Seifert, Hans Helge; Franke, Knut H.; Oya, Mototsugu; Ackermann, Rolf.

In: Anticancer Research, Vol. 23, No. 5 A, 09.2003, p. 3991-3999.

Research output: Contribution to journalArticle

Schmidt, Bettina ; Anastasiadis, Aristotelis G. ; Seifert, Hans Helge ; Franke, Knut H. ; Oya, Mototsugu ; Ackermann, Rolf. / Detection of Circulating Prostate Cells during Radical Prostatectomy by Standardized PSMA RT-PCR : Association with Positive Lymph Nodes and High Malignant Grade. In: Anticancer Research. 2003 ; Vol. 23, No. 5 A. pp. 3991-3999.
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abstract = "Background: The purpose of this study was to establish a quantitative standardized RT-PCR for the detection of Prostate Specific Membrane Antigen (PSMA)-expressing circulating cells and to evaluate clinical relevance in patients undergoing radical retropubic prostatectomy (RRP) for clinically localized prostate cancer (PCa). Materials and Methods: An external standard molecule (PSMA MIMIC) was constructed for standardization of PSMA RT-PCR reactions. It has the same sequence as endogenous PSMA, except for a central 85 bp deletion, allowing the amplification of both targets simultaneously with nearly the same amplification characteristics as in a nested PCR. PSMA RT-PCR was performed from peripheral blood samples of 73 patients with clinically localized PCa, 4 with metastatic PCa, 27 with benign prostatic hyperplasia (BPH) and 27 controls. Pre, intra- and postoperative blood samples were tested for circulating PSMA expressing cells in 54 out of 73 patients with clinically localized PCa. We also tested intraoperative blood samples of 19 BPH patients treated by transurethral or open surgery. Results: Endogenous PSMA signals from PCa patients varied between 850 and 9900 transcript molecules, corresponding to 2-20 PSMA-expressing cells/ml blood. Standardized RT-PCR using the PSMA MIMIC molecule revealed a significant decrease of {"}false-positives{"} in cancer-free controls (p=0.004). Controls could clearly be distinguished from prostate cancer patients based on PSMA PCR positivity (p=0.003). Thirty-two {\%} of patients with localized prostate cancer, 11{\%} of BPH patients and 7{\%} of healthy controls were positive in standardized assays compared to 48{\%}, 30{\%} and 27{\%} without PSMA MIMIC, respectively. Preoperatively, a correlation with tumor stage (p=0.030), grade (p=0.035) and Gleason Score (p=0.03) could be demonstrated in clinically localized PCa patients. Dissemination of prostate cells during surgery occurred in 32{\%} of the RRPs and 21{\%} of BPH patients. Positive PCR signals from intraoperative blood samples correlated with positive lymph node status (p=0.007) and tumor grade (p=0.005). Postoperative positive results correlated with grade (p=0.012) and Gleason Score (p=0.035). Conclusion: Counseling the patient with clinically localized prostate cancer can be challenging. Surgery may be, in retrospect, inappropriate in a number of patients due to preoperative understaging. This newly constructed external standard allows quantitative detection of circulating prostate cells, and therefore may open new perspectives for PSMA RT-PCR techniques as diagnostic assays and tools for post-therapeutic follow-up.",
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T1 - Detection of Circulating Prostate Cells during Radical Prostatectomy by Standardized PSMA RT-PCR

T2 - Association with Positive Lymph Nodes and High Malignant Grade

AU - Schmidt, Bettina

AU - Anastasiadis, Aristotelis G.

AU - Seifert, Hans Helge

AU - Franke, Knut H.

AU - Oya, Mototsugu

AU - Ackermann, Rolf

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N2 - Background: The purpose of this study was to establish a quantitative standardized RT-PCR for the detection of Prostate Specific Membrane Antigen (PSMA)-expressing circulating cells and to evaluate clinical relevance in patients undergoing radical retropubic prostatectomy (RRP) for clinically localized prostate cancer (PCa). Materials and Methods: An external standard molecule (PSMA MIMIC) was constructed for standardization of PSMA RT-PCR reactions. It has the same sequence as endogenous PSMA, except for a central 85 bp deletion, allowing the amplification of both targets simultaneously with nearly the same amplification characteristics as in a nested PCR. PSMA RT-PCR was performed from peripheral blood samples of 73 patients with clinically localized PCa, 4 with metastatic PCa, 27 with benign prostatic hyperplasia (BPH) and 27 controls. Pre, intra- and postoperative blood samples were tested for circulating PSMA expressing cells in 54 out of 73 patients with clinically localized PCa. We also tested intraoperative blood samples of 19 BPH patients treated by transurethral or open surgery. Results: Endogenous PSMA signals from PCa patients varied between 850 and 9900 transcript molecules, corresponding to 2-20 PSMA-expressing cells/ml blood. Standardized RT-PCR using the PSMA MIMIC molecule revealed a significant decrease of "false-positives" in cancer-free controls (p=0.004). Controls could clearly be distinguished from prostate cancer patients based on PSMA PCR positivity (p=0.003). Thirty-two % of patients with localized prostate cancer, 11% of BPH patients and 7% of healthy controls were positive in standardized assays compared to 48%, 30% and 27% without PSMA MIMIC, respectively. Preoperatively, a correlation with tumor stage (p=0.030), grade (p=0.035) and Gleason Score (p=0.03) could be demonstrated in clinically localized PCa patients. Dissemination of prostate cells during surgery occurred in 32% of the RRPs and 21% of BPH patients. Positive PCR signals from intraoperative blood samples correlated with positive lymph node status (p=0.007) and tumor grade (p=0.005). Postoperative positive results correlated with grade (p=0.012) and Gleason Score (p=0.035). Conclusion: Counseling the patient with clinically localized prostate cancer can be challenging. Surgery may be, in retrospect, inappropriate in a number of patients due to preoperative understaging. This newly constructed external standard allows quantitative detection of circulating prostate cells, and therefore may open new perspectives for PSMA RT-PCR techniques as diagnostic assays and tools for post-therapeutic follow-up.

AB - Background: The purpose of this study was to establish a quantitative standardized RT-PCR for the detection of Prostate Specific Membrane Antigen (PSMA)-expressing circulating cells and to evaluate clinical relevance in patients undergoing radical retropubic prostatectomy (RRP) for clinically localized prostate cancer (PCa). Materials and Methods: An external standard molecule (PSMA MIMIC) was constructed for standardization of PSMA RT-PCR reactions. It has the same sequence as endogenous PSMA, except for a central 85 bp deletion, allowing the amplification of both targets simultaneously with nearly the same amplification characteristics as in a nested PCR. PSMA RT-PCR was performed from peripheral blood samples of 73 patients with clinically localized PCa, 4 with metastatic PCa, 27 with benign prostatic hyperplasia (BPH) and 27 controls. Pre, intra- and postoperative blood samples were tested for circulating PSMA expressing cells in 54 out of 73 patients with clinically localized PCa. We also tested intraoperative blood samples of 19 BPH patients treated by transurethral or open surgery. Results: Endogenous PSMA signals from PCa patients varied between 850 and 9900 transcript molecules, corresponding to 2-20 PSMA-expressing cells/ml blood. Standardized RT-PCR using the PSMA MIMIC molecule revealed a significant decrease of "false-positives" in cancer-free controls (p=0.004). Controls could clearly be distinguished from prostate cancer patients based on PSMA PCR positivity (p=0.003). Thirty-two % of patients with localized prostate cancer, 11% of BPH patients and 7% of healthy controls were positive in standardized assays compared to 48%, 30% and 27% without PSMA MIMIC, respectively. Preoperatively, a correlation with tumor stage (p=0.030), grade (p=0.035) and Gleason Score (p=0.03) could be demonstrated in clinically localized PCa patients. Dissemination of prostate cells during surgery occurred in 32% of the RRPs and 21% of BPH patients. Positive PCR signals from intraoperative blood samples correlated with positive lymph node status (p=0.007) and tumor grade (p=0.005). Postoperative positive results correlated with grade (p=0.012) and Gleason Score (p=0.035). Conclusion: Counseling the patient with clinically localized prostate cancer can be challenging. Surgery may be, in retrospect, inappropriate in a number of patients due to preoperative understaging. This newly constructed external standard allows quantitative detection of circulating prostate cells, and therefore may open new perspectives for PSMA RT-PCR techniques as diagnostic assays and tools for post-therapeutic follow-up.

KW - Prostate cancer

KW - PSMA

KW - Radical prostatectomy

KW - RT-PCR

KW - Standardization

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